Molecular cloning, spatial and temporal expression analysis of CatSper genes in the Chinese Meishan pigs

Table 2 Porcine primer sequences and PCR protocols for the genes used in RT-PCR analysis.

Primers	Sequence 5'-3'		Product size (bp)	PCR protocol
	Forward	Reverse		94°C	94°C	60°C	72°C	72°C
CatSper1 ^a	gagcatcctcaccaccatct	gtgttcgctcatcacctcct	310	3 min	30 sec	30 sec	30 sec	5 min
	ggtgactcccacctctttga	cttcttttctggctcgatgg	111	3 min	30 sec	30 sec	20 sec	5 min
CatSper2 ^a	tgttttcatcttggagatccttc	gtagacaccagccacagcaa	319	3 min	30 sec	30 sec	30 sec	5 min
	cttgcacgattccctcaaat	gcgagttgaacgggtgtaat	153	3 min	30 sec	30 sec	20 sec	5 min
CatSper3 ^a	ctactggagggacggctaca	gccaagctgaagagggtaaa	348	3 min	30 sec	30 sec	40 sec	5 min
	gctggtgcagacacagaaaa	atcctggttgtccagagtgg	160	3 min	30 sec	30 sec	20 sec	5 min
CatSper4 ^a	acctctcggaaacacatgc	ggctgggtgtgtttgcttat	350	3 min	30 sec	30 sec	35 sec	5 min
	ggacacccgtgagatgactt	ggctgggtgtgtttgcttat	157	3 min	30 sec	30 sec	20 sec	5 min
GAPDH ^a	acccagaagactgtggatgg	ccccagcatcaaaggtagaa	346	3 min	30 sec	30 sec	35 sec	5 min
	catggcctccaaggagtaag	tctgggatggaaactggaag	150	3 min	30 sec	30 sec	20 sec	5 min
CD4	tggaaacctgaccttggttc	agaacccagcgagaaacaga	359	3 min	30 sec	30 sec	35 sec	5 min
E-Cadherin	ccccatgttcgaatatccac	tgctgtgacaagccatcttc	396	3 min	30 sec	30 sec	40 sec	5 min
c-kit	ttgttgatgacctcgtggaa	atggaatctgaggccttcct	307	3 min	30 sec	30 sec	30 sec	5 min

^aFirst primer pairs were used for the qualitative RT-PCR, and second primer pairs were used for the quantitative real-time RT-PCR.

ISSN: 1477-7827